Method and apparatus for examining diseases with inborn errors of metabolism

A detection method and technology of detection equipment, which are applied to the determination/inspection of microorganisms, biochemical equipment and methods, measuring devices, etc., can solve the problems of convenience and speed that need to be improved, avoid the need for preparing reagents, and simplify the operation. Effect

Inactive Publication Date: 2003-12-10
SAPPORO IMMUNO DIAGNOSTIC LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, if the typical point-of-care testing (POCT) of blood glucose monitoring in diab

Method used

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  • Method and apparatus for examining diseases with inborn errors of metabolism
  • Method and apparatus for examining diseases with inborn errors of metabolism
  • Method and apparatus for examining diseases with inborn errors of metabolism

Examples

Experimental program
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Embodiment 1

[0066] Examples of the present invention will now be specifically described, but the present invention is not limited to these examples. The preparation of embodiment 1 sensor chip

[0067] attached figure 1 An exploded perspective view showing an example of fabricating a sensor chip and is an exploded view of the main components. The sensor chip consists of a transparent window 15 for optical detection, an electrode system formed by screen printing, a layered structure 6 with a reaction reagent immobilized inside, a spacer 10 and a cover plate 11 .

[0068] In forming the electrode system, conductive silver powder ink (Nippon Acheson) was screen-printed on the insulating plate 1 of PET film (TORAY INDUSTRIES) and then dried by heating (120° C., 15 minutes) to form wires 2 . Using insulating ink (Nippon Acheson) an insulating layer 3 is then made (cured by UV irradiation), which part is connected to the working electrode 4 and the counter electrode 5 held below it. Working ...

Embodiment 2

[0072] A reaction reagent immobilization layer 6 of a layered structure in which a desired reaction reagent is immobilized is placed so as to be sandwiched between backing plates 10 on electrodes formed by printing. This composite is covered with a cover plate 11 in order to produce a sensor chip 17 . The basic response of embodiment 2 sensor chip

[0073] Special sensor chips for GE, MSUD and PKU detection were prepared according to the manufacturing steps in Example 1. Only dehydrogenases specific for D-Gal, branched-chain amino acids including L-Leu, and L-Phe as substrates were changed and all other reaction reagents were the same.

[0074] For the GE detection sensor chip, D-Gal dehydrogenase (EC 1.1.1.48, Roche Diagnostics) was used.

[0075] Phosphorylated Gal (galactose 1-phosphate) and Gal can be determined by the presence of alkaline phosphatase (EC 3.1.3.1).

[0076] For the MSUD detection sensor chip, L-Leu dehydrogenase (EC 1.4.1.9, TOYOBO) was used. For the P...

Embodiment 3

[0081] In the concentration range up to 2 mM, both detection methods and corresponding sensor chips yield corresponding values ​​that are dependent on the substrate concentration. The production of embodiment 3 simple counters

[0082] The simple counter of the assay device using GE, MSUD and PKU detection sensor chips is attached Figure 4 , and the detection process is explained as follows.

[0083] Each sensor chip 17 is mounted on a simple counter 18 through a chip insertion portion 19 . At this time, there is no restriction on the start of the counter; the counter can be started by the switch 21 or the counter can be started automatically.

[0084] The sample 13 is dropped on the sensor chip 17 and measurement is started. Also in this case, there is no restriction as to whether the measurement is performed by the switch 21 or is started automatically.

[0085] The generated enzymatic reaction and oxidation-reduction reaction are performed for a certain period of time ...

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Abstract

A method of conveniently and quickly quantitating subject substances which comprises using as reaction reagents at least dehydrogenases specific respectively to branched amino acids containing D-galactose and L-leucine and phenylalanine, which are the substances contained in biological samples to be assayed, coenzymes, an electron mediator and a tetrazolium salt and detecting formazan, which has been finally formed depending on the concentrations of the subject substances by the enzymatic reactions and oxidation reactions of the biological samples with the reaction reagents, by using both or one of an optical detection procedure and an electrochemical detection procedure, thereby embodying a convenient and quick examination of three diseases with inborn errors of metabolism, i.e., galactosemia, maple syrup urine disease and phenylketonuria.

Description

field of invention [0001] The present invention relates to a method for measuring D-galactose, branched-chain amino acids including L-leucine, and L-phenylalanine which are contained in biological samples simply and rapidly without requiring troublesome pretreatment and Determination equipment. [0002] Especially in the three diseases of inborn errors of metabolism, galactosemia, maple diabetes and phenylketonuria, the method and device can be used to screen newborns for early detection of these diseases. Bedside examination and daily monitoring of patients with these disorders. Background of the invention [0003] Worldwide newborn screening for early detection of inborn errors of metabolism began with treatment for phenylketonuria (PKU) and for L-phenylalanine (L-Phe ) The discovery of a semi-quantitative determination method. [0004] The main reason why inborn errors of metabolism become target diseases for screening is the elucidation of medical advantages, making i...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N33/66G01N33/68
CPCG01N33/52G01N33/6812G01N33/66
Inventor 横山撤筱塚直树中村健治
Owner SAPPORO IMMUNO DIAGNOSTIC LAB
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